inflammatory responses and immune cell populations in skin explants - PowerPoint PPT Presentation

PUFA-derived bioactive lipids mediate inflammatory responses and immune cell populations in skin explants Alexandra Kendall Nordic LipidForum 6 th June 2015

SLS vs UVR SLS UVR Commonly encountered environmental stressors • Exposure to high levels of SLS can causes irritant dermatitis • - Dermatology/occupational health problem • UVR is an alternative cause of skin erythema • Do they affect the same lipid pathways?

Ex vivo culture

Ex vivo treatment Day 1 (evening): Delivery and processing of skin Day 2 (morning): Culture medium replaced Day 3 (morning): SDS-soaked disc Culture medium applied replaced SDS-soaked disc Day 4 (morning): UVR applied removed Day 5 (morning): Biopsies taken from centre of pieces of baseline and UV- and SLS-treated skin

Lipoxygenase products 12-lipoxygenase 15-lipoxygenase 12-HETE 15-HETE AA 11-HDHA DHA 17 - HDHA DGLA EPA 15-HETrE 14-HDHA LA 12-HEPE 13-HODE

SLS 12-LOX products 100000 Control * * * 1% SDS 10000 pg/mg protein 3% SDS 6% SDS 1000 * * * * * 100 10 1 12 HETE 12 HEPE 14 HDHA 11 HDHA 15-LOX products * * 10000 * * Control * * * * 1% SDS 1000 pg/mg protein 3% SDS 6% SDS 100 10 1 n=4, mean ± SE 15 HETE 15 HETrE 13 HODE 17 HDHA * p<0.05

UVR 12-LOX products 10000 Control 4MED 1000 pg/mg protein 6MED 10MED 100 10 1 12 HETE 12 HEPE 14 HDHA 11 HDHA 15-LOX products 1000 Control 4MED pg/mg protein 6MED 100 10MED 10 1 15 HETE 15 HETrE 13 HODE 17 HDHA n=4, mean ± SE

N-acyl ethanolamides

SLS N-acyl ethanolamides 100000 Control * * 1% SLS * * 10000 3% SLS pg/mg protein 6% SLS 1000 * 100 10 1 PEA ALEA LEA OEA STEA EPEA AEA DHEA DGLEA n=4, mean ± SE * p<0.05

UVR N-acyl ethanolamides 10000 Control 4MED 6MED 1000 pg/mg protein 10MED 100 10 1 PEA ALEA LEA OEA STEA EPEA AEA DHEA DGLEA n=4, mean ± SE

Immune cell population identification Antigen presenting cells CD14 CD1a CD45 SSC SSC HLA-DR Innate lymphoid cells T cells Granulocytes CD66b CD127 TCRgd CD3 NK cells CD56 SSC SSC SSC

Immune cell populations HLA-DR+ HLA-DR+ T cells T cells Granulocytes Granulocytes NK cells NK cells ILCs ILCs HLA-DR+ HLA-DR+ T cells T cells Granulocytes Granulocytes NK cells NK cells ILCs ILCs • Inter-individual variation in response • Consistent effect was reduction in innate lymphoid cell population

Conclusions Despite similar clinical changes in vivo , SLS and UVR elicit different effects in skin explants SLS UVR 12-LOX products Increase Non-significant increase 15-LOX products Increase No change N-acyl ethanolamides Increase No change Epidermal innate Decrease No change lymphoid cell population

Acknowledgements University of Manchester Prof Anna Nicolaou Dr Amy Saunders Unilever SEAC Gary Sassano This work was funded by Unilever as part of its ongoing program developing novel approaches for assuring consumer safety